10.1371/journal.ppat.1007233.g005
Hongbo Guo
Hongbo
Guo
Huib Rabouw
Huib
Rabouw
Anne Slomp
Anne
Slomp
Meiling Dai
Meiling
Dai
Floor van der Vegt
Floor
van der Vegt
Jan W. M. van Lent
Jan W. M.
van Lent
Ryan McBride
Ryan
McBride
James C. Paulson
James C.
Paulson
Raoul J. de Groot
Raoul
J. de Groot
Frank J. M. van Kuppeveld
Frank
J. M. van Kuppeveld
Erik de Vries
Erik
de Vries
Cornelis A. M. de Haan
Cornelis
A. M. de Haan
Contribution of NA to virion binding signal.
Public Library of Science
2018
epithelial host cells
vivo IAV particles
endpoint binding assays
receptor-containing surfaces Interactions
equilibrium binding models
biolayer interferometric analysis
mucus layer
Multiple low-affinity HA-SIA interactions
mucus decoy receptors
Quantitative BLI analysis
NA activity
virus
epithelial cell surfaces
2018-08-13 17:39:27
Figure
https://plos.figshare.com/articles/figure/Contribution_of_NA_to_virion_binding_signal_/6962330
<p>(A-B) NA activity-dependent virus self-elution was examined for the first seconds (for PR8<sub>MtSIN</sub>; panel A) or minutes (for WSN<sub>HAMtSIN</sub>; panel B) of the dissociation phase. Viruses were loaded at 100 pM concentration to 3’SLN (PR8<sub>MtSIN</sub>) or 3’N Fetuin (WSN<sub>HAMtSIN</sub>) for 30 minutes in presence of 10 μM OC after which the sensors were washed in PBS and dissociation was examined at a range of OC concentrations as indicated in the panels. (C-F) PR8<sub>MtSIN</sub> (C, E) and WSN<sub>HAMtSIN</sub> (D, F), carrying the same HA<sub>MtSIN</sub> but either NA<sub>MtSIN</sub> (high NA activity) or NA<sub>WSN</sub> (low NA activity), respectively, were bound at 100 pM concentration to biotinylated 3’SLN (C, D) or Fc-tagged 3’N Fetuin (E, F) loaded to maximum level. Binding was performed in absence (red lines) or in the presence of a range of OC concentrations as indicated in the panels. Significant differences between initial binding curves shown in panels C-F were analyzed by IBM SPSS statistic 24. In panel C, there was no significant difference between curves with 10μM OC and 625nM OC (P>0.1), whereas the curves of 10μM OC and 625nM OC were significantly different from 39nM, 2.45nM and 0nM (P<0.001). In panel D, the curves of 10μM OC and 625nM were significantly different from those of 39nM, 2.45nM and 0nM. There also was a significant difference between 39nM and 2.45nM. In panel E, the 100nM curves significantly differed from the other three OC concentrations. In panel F, there were significant differences between the 100nM (and 25nM) and 6.25nM and 0nM curves. (G) The ratio (initial virus binding rate <i>v</i><sup><i>obs</i></sup> in absence of OC)/(initial virus binding rate <i>v</i><sup><i>obs</i></sup> in presence of 10 μM OC) of four viruses carrying the WSN<sub>WT</sub> NA in the background four different HAs was plotted (the individual binding curves are shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1007233#ppat.1007233.s009" target="_blank">S9 Fig</a>). Standard deviations and significant differences between the mean initial binding rate ratios are indicated (* indicates P<0.05).</p>